5-Benzylidene-4-Oxazolidinones

ABSTRACT

Provided herein are compounds that can exhibit activity as biofilm modulating agents (e.g., activity as biofilm inhibitors and/or activity as biofilm dispersal agents). The compounds can exhibit potent activity against Gram positive biofilms. The compounds can also exhibit activity against Gram negative biofilms. In some cases, the compounds can exhibit both biofilm modulation properties and antimicrobial activity. Compositions comprising these compounds, as well as methods of using thereof, are also described. For example, the compounds described herein can be used in human and animal health (e.g., for the treatment of infection), agriculture, marine coatings, and other coating applications related to prevention of biofilm (e.g., dental, medical, etc.).

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with Government Support under Grant No. GM110154 awarded by the National Institutes of Health. The Government hascertain rights in the invention.

BACKGROUND

Biofilms are bacterial communities encased in a hydrated extracellularmatrix, which can include proteins, polysaccharides, and/or nucleicacids. The development of biofilms on biological and inanimate surfacespresents significant medical problems. Bacteria in the biofilm mode ofgrowth are highly resistant to treatment with antibiotics and toclearance by a host's immune system. Therefore, once these bacterialcommunities form, they are extremely difficult to eradicate withconventional treatments. Hence, biofilms can lead to chronic systemicinfections. For example, bacterial biofilms have been found in humanpatients associated with a variety of diseases, including, urinary tractinfections, middle ear infections, dental plaque, gingivitis,endocarditis, and the respiratory tract of cystic fibrosis patients.Pathogenic bacteria may form biofilms on a variety of medical implantsas well, such as indwelling catheters, artificial heart valves, andpacemakers.

The most clinically relevant characteristic of biofilm bacteria is thatthey are up to 1000-fold more resistant to antibiotics and biocides thanare planktonic bacteria. In addition, biofilm bacteria have alsodemonstrated resistance to phagocytosis by sentinel leukocytes of theimmune system. Accordingly, biofilm bacteria can survive conventionalantibiotic treatments, evade a host's immune system, and provide areservoir of infectious bacteria that can cause recurrent chronicinfections.

Biofilm-related infections are currently treated with antibiotics orantibiotic combinations that are optimized to treat infections caused byplanktonic bacteria. These treatments usually resolve the symptoms ofinfection by killing the planktonic bacteria released from the biofilm.However, these existing treatments are generally ineffective against theunderlying biofilms associated with the infection.

Accordingly, there is a critical need for compounds and compositionsthat can control biofilms, as well as improved methods for controllingbiofilms.

SUMMARY

Provided herein are compounds that can exhibit activity as biofilmmodulating agents (e.g., activity as biofilm inhibitors and/or activityas biofilm dispersal agents). The compounds can exhibit potent activityagainst Gram positive biofilms. The compounds can also exhibit activityagainst Gram negative biofilms. In some cases, the compounds can exhibitboth biofilm modulation properties and antimicrobial activity.

For example, provided herein are compounds defined by Formula I

or a pharmaceutically acceptable salt or prodrug thereof, wherein X ischosen from O and S; R¹, R², R³, R⁴, and R⁵ are each independentlychosen from hydrogen, halogen, hydroxyl, —CN, —NO₂, amino, alkylamino,dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy,haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl,haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl,haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁶ and R⁷ are each independently chosen fromhydrogen, hydroxy, halogen, —CN, —NO₂, amino, alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl, haloalkylsulfinyl,alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl,alkoxycarbonyl, haloalkoxycarbonyl, alkylaminocarbonyl,heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁸ is chosen from alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, alkylcycloheteroalkyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl, each optionallysubstituted with one or more substituents individually chosen from R¹¹;R⁹ is chosen from hydrogen, hydroxy, halogen, —CN, —NO₂, amino,alkylamino, dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio,alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, aryl,heteroaryl, cycloalkyl, cycloheteroalkyl, alkylaryl, alkylheteroaryl,alkylcycloalkyl, alkylcycloheteroalkyl, alkylsulfinyl,haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl,haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl, each optionally substituted with one or moresubstituents individually chosen from R¹¹; R¹⁰ is chosen from hydrogen,alkyl, haloalkyl, alkylthio, haloalkylthio, alkenyl, haloalkenyl,alkynyl, haloalkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, and alkylcycloheteroalkyl,each optionally substituted with one or more substituents individuallychosen from R¹¹; and R¹¹ is chosen from hydroxy, halogen, —CN, —NO₂,amino, alkylamino, dialkylamino, alkyl, haloalkyl; alkylthio;haloalkylthio; alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl. In someembodiments, the compound is not one of the following:

In some embodiments, X can be O.

In some embodiments, at least one of R¹, R², R³, R⁴, and R⁵ is nothydrogen. In some cases, R¹, R², R⁴, and R⁵ are hydrogen. In certaincases, R¹, R², R⁴, and R⁵ are hydrogen, and R⁴ is not hydrogen (e.g.,the phenyl ring of the benzylidene moiety is para-substituted).

In some embodiment, R³ can be an electron withdrawing group. Forexample, R³ can be chosen from halogen, —CN, —NO₂, haloalkyl,alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl, andhaloalkoxycarbonyl. In some embodiments, R³ can be a haloalkyl group. Incertain embodiments, R³ can be a perfluoroalkyl group (e.g., a —CF₃group).

In some embodiments, at least one of R⁶ and R⁷ is not hydrogen. Incertain embodiments, both R⁶ and R⁷ are not hydrogen. In some cases, atleast one of R⁶ and R⁷ can be halogen (e.g., at least one of R⁶ and R⁷can be —Cl and/or at least one of R⁶ and R⁷ can be —F).

In some cases, both R⁶ and R⁷ can be halogen (e.g., both R⁶ and R⁷ canbe —Cl, or both R⁶ and R⁷ can be —F).

In some examples, R⁸ can comprise a substituent having from 5 to 12carbon atoms. In certain examples, R⁸ can be chosen from alkyl, aryl,heteroaryl, alkylaryl, and alkylheteroaryl. In some embodiments, R⁸ cancomprise a C₁-C₈ alkyl group (e.g., a C₁-C₄ alkyl group, or a C₅-C₈alkyl group). In other embodiments, R⁸ can comprise a C₆-C₁₀ alkylarylgroup (e.g., a substituted or unsubstituted benzyl group).

In some embodiments, R⁹ can be hydrogen. In other embodiments, R⁹ cancomprise an alkyl group (e.g., a C₁-C₄ alkyl group) or an aryl group(e.g., a phenyl group).

In some embodiments, R¹⁰ can be hydrogen. In other embodiments, R¹⁰ canbe an alkyl group, an aryl group, or an alkylaryl group. In certainembodiments, R10 can comprise an alkyl group (e.g., a C₁-C₄ alkyl group)or an aryl group (e.g., a phenyl group).

In some cases, the compound can be a compound defined by Formula II

or a pharmaceutically acceptable salt or prodrug thereof, wherein R³comprises an electron withdrawing group; R⁶ and R⁷ are eachindependently chosen from hydrogen and halogen, with the proviso that atleast one of R⁶ and R⁷ is halogen; R⁸ is chosen from alkyl, haloalkyl,alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, aryl,heteroaryl, cycloalkyl, cycloheteroalkyl, alkylaryl, alkylheteroaryl,alkylcycloalkyl, alkylcycloheteroalkyl, each optionally substituted withone or more substituents individually chosen from R¹¹; and R¹¹ is chosenfrom hydroxy, halogen, —CN, —NO₂, amino, alkylamino, dialkylamino,alkyl, haloalkyl; alkylthio; haloalkylthio; alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl, haloalkylsulfinyl,alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl,alkoxycarbonyl, haloalkoxycarbonyl, alkylaminocarbonyl,heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl. In some embodiments, the compound is not oneof the following:

In some embodiments, R³ can be chosen from halogen, —CN, —NO₂,haloalkyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl, andhaloalkoxycarbonyl. In some embodiments, R³ can be a haloalkyl group. Incertain embodiments, R³ can be a perfluoroalkyl group (e.g., a —CF₃group).

Also provided are compositions that can prevent, remove, and/or inhibitbiofilms. Biofilm preventing, removing, or inhibiting compositions cancomprise a carrier and an effective amount of a compound describedherein to prevent, remove, and/or inhibit a biofilm. The composition canbe, for example, a dentifrice composition (e.g., a toothpaste,mouthwash, chewing gum, dental floss, or dental cream) that promotesdental hygiene by preventing, reducing, inhibiting or removing abiofilm.

Also provided herein are pharmaceutical compositions that comprise acompound described herein in a pharmaceutically acceptable carrier. Insome embodiments, pharmaceutical compositions can further include one ormore additional active agents (e.g., one or more antibiotics).

The compounds described herein can also be disposed on or within asubstrate to control biofilm formation on the substrate. Accordingly,also provided are medical devices that comprise a medical devicesubstrate and an effective amount of a compound described herein eithercoating the substrate, or incorporated into the substrate. The effectiveamount of the compound can be an effective amount to prevent or inhibitgrowth of a biofilm on the medical device substrate. The medical devicesubstrate can include, for example, a stent, fastener, port, catheter,scaffold, and/or graft.

Also provided herein are methods for controlling biofilm formation on asubstrate. Methods for controlling biofilm formation on a substrate cancomprise contacting the substrate with a compound described herein in anamount effective to inhibit biofilm formation. The biofilm can compriseGram-positive bacteria or Gram-negative bacteria. In some embodiments,the biofilm can comprise Gram-positive bacteria (e.g., a bacteria of agenus Staphylococcus, such as Staphylococcus aureus).

Also provided herein are methods for treating chronic bacterialinfections. Methods for treating a chronic bacterial infection in asubject in need thereof can comprise administering to said subject acompound described herein in an amount effective to inhibit, reduce, orremove a biofilm component of the chronic bacterial infection. Thechronic bacterial infection can comprise, for example, a urinary tractinfection, gastritis, a respiratory infection, cystitis, pyelonephritis,osteomyelitis, bacteremia, a skin infection, rosacea, acne, a chronicwound infection, infectious kidney stones, bacterial endocarditis, or asinus infection.

Also provided are methods of treating subjects infected with abacterium. Methods of treating a subject infected with a bacterium cancomprise administering to the subject a therapeutically effective amountof a compound described herein. In some embodiments, the bacterium cancomprise a Gram-positive bacterium. For example, the bacterium caninclude Staphylococcus aureus (methicillin sensitive), Staphylococcusaureus (methicillin resistant), Streptococcus pneumonia (penicillinsensitive), Streptococcus pneumonia (penicillin resistant),Staphylococcus epidermis (multiple drug resistant), Enterococcusfaecalis (vancomycin sensitive), Enterococcus faecium (vancomycinresistant), and/or Haemophilus influenzae.

DETAILED DESCRIPTION Definitions

Terms used herein will have their customary meaning in the art unlessspecified otherwise. The organic moieties mentioned when definingvariable positions within the general formulae described herein (e.g.,the term “halogen”) are collective terms for the individual substituentsencompassed by the organic moiety. The prefix Cn-Cm indicates in eachcase the possible number of carbon atoms in the group.

As used herein, by a “subject” is meant an individual. Thus, the“subject” can include domesticated animals (e.g., cats, dogs, etc.),livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), laboratoryanimals (e.g., mouse, rabbit, rat, guinea pig, etc.), and birds.“Subject” can also include a mammal, such as a primate or a human.

By “reduce” or other forms of the word, such as “reducing” or“reduction,” is meant lowering of an event or characteristic (e.g.,biofilm growth). It is understood that this is typically in relation tosome standard or expected value, in other words it is relative, but thatit is not always necessary for the standard or relative value to bereferred to. For example, “reducing the biofilm component of a chronicbacterial infection” can refer to reducing the rate of growth of abiofilm component of the chronic bacterial infection relative to astandard or a control.

By “prevent” or other forms of the word, such as “preventing” or“prevention,” is meant to stop a particular event or characteristic, tostabilize or delay the development or progression of a particular eventor characteristic, or to minimize the chances that a particular event orcharacteristic will occur. Prevent does not require comparison to acontrol as it is typically more absolute than, for example, reduce. Asused herein, something could be reduced but not prevented, but somethingthat is reduced could also be prevented. Likewise, something could beprevented but not reduced, but something that is prevented could also bereduced. It is understood that where reduce or prevent are used, unlessspecifically indicated otherwise, the use of the other word is alsoexpressly disclosed.

By “treat” or other forms of the word, such as “treated” or “treatment,”is meant to administer a composition or to perform a method in order toreduce, prevent, inhibit, or eliminate a particular characteristic orevent (e.g., a biofilm). The term “control” is used synonymously withthe terms “treat” and “modulate.”

“Biofilm” or “biofilms” refer to communities of microorganisms that areattached to a substrate. The microorganisms often excrete a protectiveand adhesive matrix of polymeric compounds. They often have structuralheterogeneity, genetic diversity, and complex community interactions.“Biofilm preventing”, “biofilm removing”, “biofilm inhibiting”, “biofilmreducing”, “biofilm resistant”, “biofilm controlling” or “antifouling”refer to prevention of biofilm formation, inhibition of theestablishment or growth of a biofilm, or decrease in the amount oforganisms that attach and/or grow upon a substrate, up to and includingthe complete removal of the biofilm.

As used herein, a “substrate” can include any living or nonlivingstructure. For example, biofilms often grow on synthetic materialssubmerged in an aqueous solution or exposed to humid air, but they alsocan form as floating mats on a liquid surface, in which case themicroorganisms are adhering to each other or to the adhesive matrixcharacteristic of a biofilm.

An “effective amount” of a biofilm preventing, removing or inhibitingcomposition is that amount which is necessary to carry out thecomposition's function of preventing, removing or inhibiting a biofilm.

The term “alkyl,” as used herein, refers to saturated straight,branched, cyclic, primary, secondary or tertiary hydrocarbons, includingthose having 1 to 20 atoms. In some embodiments, alkyl groups willinclude C₁-C₁₂, C₁-C₁₀, C₁-C₅, C₁-C₆, C₁-C₅, C₁-C₄, C₁-C₃, C₁-C₂, or C₁alkyl groups. Examples of C₁-C₁₀ alkyl groups include, but are notlimited to, methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl,2-methylpropyl, 1,1-dimethylethyl, pentyl, 1-methylbutyl, 2-methylbutyl,3-methylbutyl, 2,2-dimethylpropyl, 1-ethylpropyl, hexyl,1,1-dimethylpropyl, 1,2-dimethylpropyl, 1-methylpentyl, 2-methylpentyl,3-methylpentyl, 4-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl,1,3-dimethylbutyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl,3,3-dimethylbutyl, 1-ethylbutyl, 2-ethylbutyl, 1,1,2-trimethylpropyl,1,2,2-trimethylpropyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl,heptyl, octyl, 2-ethylhexyl, nonyl and decyl groups, as well as theirisomers. Examples of C₁-C₄-alkyl groups include, for example, methyl,ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl, 2-methylpropyl and1,1-dimethylethyl groups.

Cyclic alkyl groups or “cycloalkyl” groups, which are encompassed alkyl,include cycloalkyl groups having from 3 to 10 carbon atoms. Cycloalkylgroups can include a single ring, or multiple condensed rings. In someembodiments, cycloalkyl groups include C₃-C₄, C₄-C₇, C₅-C₇, C₄-C₆, orC₅-C₆ cyclic alkyl groups. Non-limiting examples of cycloalkyl groupsinclude adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl and the like.

Alkyl groups can be unsubstituted or substituted with one or moremoieties selected from the group consisting of alkyl, halo, haloalkyl,hydroxyl, carboxyl, acyl, acyloxy, amino, alkyl- or dialkylamino, amido,arylamino, alkoxy, aryloxy, nitro, cyano, azido, thiol, imino, sulfonicacid, sulfate, sulfonyl, sulfanyl, sulfinyl, sulfamonyl, ester,phosphonyl, phosphinyl, phosphoryl, phosphine, thioester, thioether,acid halide, anhydride, oxime, hydrazine, carbamate, phosphoric acid,phosphate, phosphonate, or any other viable functional group that doesnot inhibit the biological activity of the compounds of the invention,either unprotected, or protected as necessary, as known to those skilledin the art, for example, as described in Greene, et al., ProtectiveGroups in Organic Synthesis, John Wiley and Sons, Third Edition, 1999,hereby incorporated by reference.

Terms including the term “alkyl,” such as “alkylcycloalkyl,”“cycloalkylalkyl,” “alkylamino,” or “dialkylamino,” will be understoodto comprise an alkyl group as defined above linked to another functionalgroup, where the group is linked to the compound through the last grouplisted, as understood by those of skill in the art.

The term “alkenyl,” as used herein, refers to both straight and branchedcarbon chains which have at least one carbon-carbon double bond. In someembodiments, alkenyl groups can include C₂-C₂₀ alkenyl groups. In otherembodiments, alkenyl can include C₂-C₁₂, C₂-C₁₀, C₂-C₈, C₂-C₆ or C₂-C₄alkenyl groups. In one embodiment of alkenyl, the number of double bondsis 1-3, in another embodiment of alkenyl, the number of double bonds isone or two. Other ranges of carbon-carbon double bonds and carbonnumbers are also contemplated depending on the location of the alkenylmoiety on the molecule. “C₂-C₁₀-alkenyl” groups may include more thanone double bond in the chain. The one or more unsaturations within thealkenyl group may be located at any position(s) within the carbon chainas valence permits. In some embodiments, when the alkenyl group iscovalently bound to one or more additional moieties, the carbon atom(s)in the alkenyl group that are covalently bound to the one or moreadditional moieties are not part of a carbon-carbon double bond withinthe alkenyl group. Examples of alkenyl groups include, but are notlimited to, ethenyl, 1-propenyl, 2-propenyl, 1-methyl-ethenyl,1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-1-propenyl,2-methyl-1-propenyl, 1-methyl-2-propenyl, 2-methyl-2-propenyl;1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl,2-methyl-1-butenyl, 3-methyl-1-butenyl, 1-methyl-2-butenyl,2-methyl-2-butenyl, 3-methyl-2-butenyl, 1-methyl-3-butenyl,2-methyl-3-butenyl, 3-methyl-3-butenyl, 1,1-dimethyl-2-propenyl,1,2-dimethyl-1-propenyl, 1,2-dimethyl-2-propenyl, 1-ethyl-1-propenyl,1-ethyl-2-propenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl,5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl,3-methyl-1-pentenyl, 4-methyl-1-pentenyl, 1-methyl-2-pentenyl,2-methyl-2-pentenyl, 3-methyl-2-pentenyl, 4-methyl-2-pentenyl,1-methyl-3-pentenyl, 2-methyl-3-pentenyl, 3-methyl-3-pentenyl,4-methyl-3-pentenyl, 1-methyl-4-pentenyl, 2-methyl-4-pentenyl,3-methyl-4-pentenyl, 4-methyl-4-pentenyl, 1,1-dimethyl-2-butenyl,1,1-dimethyl-3-butenyl, 1,2-dimethyl-1-butenyl, 1,2-dimethyl-2-butenyl,1,2-dimethyl-3-butenyl, 1,3-dimethyl-1-butenyl, 1,3-dimethyl-2-butenyl,1,3-dimethyl-3-butenyl, 2,2-dimethyl-3-butenyl, 2,3-dimethyl-1-butenyl,2,3-dimethyl-2-butenyl, 2,3-dimethyl-3-butenyl, 3,3-dimethyl-1-butenyl,3,3-dimethyl-2-butenyl, 1-ethyl-1-butenyl, 1-ethyl-2-butenyl,1-ethyl-3-butenyl, 2-ethyl-1-butenyl, 2-ethyl-2-butenyl,2-ethyl-3-butenyl, 1,1,2-trimethyl-2-propenyl,1-ethyl-1-methyl-2-propenyl, 1-ethyl-2-methyl-1-propenyl and1-ethyl-2-methyl-2-propenyl groups.

The term “alkynyl,” as used herein, refers to both straight and branchedcarbon chains which have at least one carbon-carbon triple bond. In oneembodiment of alkynyl, the number of triple bonds is 1-3; in anotherembodiment of alkynyl, the number of triple bonds is one or two. In someembodiments, alkynyl groups include from C₂-C₂₀ alkynyl groups. In otherembodiments, alkynyl groups may include C₂-C₁₂, C₂-C₁₀, C₂-C₈, C₂-C₆ orC₂-C₄ alkynyl groups. Other ranges of carbon-carbon triple bonds andcarbon numbers are also contemplated depending on the location of thealkenyl moiety on the molecule. For example, the term “C₂-C₁₀-alkynyl”as used herein refers to a straight-chain or branched unsaturatedhydrocarbon group having 2 to 10 carbon atoms and containing at leastone triple bond, such as ethynyl, prop-1-yn-1-yl, prop-2-yn-1-yl,n-but-1-yn-1-yl, n-but-1-yn-3-yl, n-but-1-yn-4-yl, n-but-2-yn-1-yl,n-pent-1-yn-1-yl, n-pent-1-yn-3-yl, n-pent-1-yn-4-yl, n-pent-1-yn-5-yl,n-pent-2-yn-1-yl, n-pent-2-yn-4-yl, n-pent-2-yn-5-yl,3-methylbut-1-yn-3-yl, 3-methylbut-1-yn-4-yl, n-hex-1-yn-1-yl,n-hex-1-yn-3-yl, n-hex-1-yn-4-yl, n-hex-1-yn-5-yl, n-hex-1-yn-6-yl,n-hex-2-yn-1-yl, n-hex-2-yn-4-yl, n-hex-2-yn-5-yl, n-hex-2-yn-6-yl,n-hex-3-yn-1-yl, n-hex-3-yn-2-yl, 3-methylpent-1-yn-1-yl,3-methylpent-1-yn-3-yl, 3-methylpent-1-yn-4-yl, 3-methylpent-1-yn-5-yl,4-methylpent-1-yn-1-yl, 4-methylpent-2-yn-4-yl, and4-methylpent-2-yn-5-yl groups.

The term “haloalkyl,” as used herein refers to an alkyl group, asdefined above, which is substituted by one or more halogen atoms. Insome instances, the haloalkyl group can be an alkyl group substituted byone or more fluorine atoms. In certain instances, the haloalkyl groupcan be a perfluorinated alkyl group. For example C₁-C₄-haloalkylincludes, but is not limited to, chloromethyl, bromomethyl,dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl,trifluoromethyl, chlorofluoromethyl, dichlorofluoromethyl,chlorodifluoromethyl, 1-chloroethyl, 1-bromoethyl, 1-fluoroethyl,2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl,2-chloro-2-fluoroethyl, 2-chloro-2,2-difluoroethyl,2,2-dichloro-2-fluoroethyl, 2,2,2-trichloroethyl, and pentafluoroethyl.

The term “haloalkenyl,” as used herein, refers to an alkenyl group, asdefined above, which is substituted by one or more halogen atoms.

The term “haloalkynyl,” as used herein, refers to an alkynyl group, asdefined above, which is substituted by one or more halogen atoms.

The term “alkoxy,” as used herein, refers to alkyl-O—, wherein alkylrefers to an alkyl group, as defined above. Similarly, the terms“alkenyloxy,” “alkynyloxy,” “haloalkoxy,” “haloalkenyloxy,”“haloalkynyloxy,” “cycloalkoxy,” “cycloalkenyloxy,” “halocycloalkoxy,”and “halocycloalkenyloxy” refer to the groups alkenyl-O—, alkynyl-O—,haloalkyl-O—, haloalkenyl-O—, haloalkynyl-O—, cycloalkyl-O—,cycloalkenyl-O—, halocycloalkyl-O—, and halocycloalkenyl-O—,respectively, wherein alkenyl, alkynyl, haloalkyl, haloalkenyl,haloalkynyl, cycloalkyl, cycloalkenyl, halocycloalkyl, andhalocycloalkenyl are as defined above. Examples of C₁-C₆-alkoxy include,but are not limited to, methoxy, ethoxy, C₂H₅—CH₂O—, (CH₃)₂CHO—,n-butoxy, C₂H₅—CH(CH₃)O—, (CH₃)₂CH—CH₂O—, (CH₃)₃CO—, n-pentoxy,1-methylbutoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy,1,2-dimethylpropoxy, 2,2-dimethyl-propoxy, 1-ethylpropoxy, n-hexoxy,1-methylpentoxy, 2-methylpentoxy, 3-methylpentoxy, 4-methylpentoxy,1,1-dimethylbutoxy, 1,2-dimethylbutoxy, 1,3-dimethylbutoxy,2,2-dimethylbutoxy, 2,3-dimethylbutoxy, 3,3-dimethylbutoxy,1-ethylbutoxy, 2-ethylbutoxy, 1,1,2-trimethylpropoxy,1,2,2-trimethylpropoxy, 1-ethyl-1-methylpropoxy, and1-ethyl-2-methylpropoxy.

The term “alkylthio,” as used herein, refers to alkyl-S—, wherein alkylrefers to an alkyl group, as defined above. Similarly, the terms“haloalkylthio,” “cycloalkylthio,” and the like, refer to haloalkyl-S—and cycloalkyl-S— where haloalkyl and cycloalkyl are as defined above.

The term “alkylsulfinyl,” as used herein, refers to alkyl-S(O)—, whereinalkyl refers to an alkyl group, as defined above. Similarly, the term“haloalkylsulfinyl” refers to haloalkyl-S(O)— where haloalkyl is asdefined above.

The term “alkylsulfonyl,” as used herein, refers to alkyl-S(O)₂—,wherein alkyl is as defined above. Similarly, the term“haloalkylsulfonyl” refers to haloalkyl-S(O)₂— where haloalkyl is asdefined above.

The terms “alkylamino” and “dialkylamino,” as used herein, refer toalkyl-NH— and (alkyl)₂N— groups, where alkyl is as defined above.Similarly, the terms “haloalkylamino” and “halodialkylamino” refer tohaloalkyl-NH— and (haloalkyl)₂-NH—, where haloalkyl is as defined above.

The terms “alkylcarbonyl,” “alkoxycarbonyl,” “alkylaminocarbonyl,” and“dialkylaminocarbonyl,” as used herein, refer to alkyl-C(O)—,alkoxy-C(O)—, alkylamino-C(O)— and dialkylamino-C(O)— respectively,where alkyl, alkoxy, alkylamino, and dialkylamino are as defined above.Similarly, the terms “haloalkylcarbonyl,” “haloalkoxycarbonyl,”“haloalkylaminocarbonyl,” and “dihaloalkylaminocarbonyl,” as usedherein, refer to the groups haloalkyl-C(O)—, haloalkoxy-C(O)—,haloalkylamino-C(O)—, and dihaloalkylamino-C(O)—, where haloalkyl,haloalkoxy, haloalkylamino, and dihaloalkylamino are as defined above.

The term “aryl,” as used herein, refers to a monovalent aromaticcarbocyclic group of from 6 to 14 carbon atoms. Aryl groups can includea single ring or multiple condensed rings. In some embodiments, arylgroups include C₆-C₁₀ aryl groups. Aryl groups include, but are notlimited to, phenyl, biphenyl, naphthyl, tetrahydronaphtyl,phenylcyclopropyl and indanyl. Aryl groups may be unsubstituted orsubstituted by one or more moieties selected from halogen, cyano, nitro,hydroxy, mercapto, amino, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, haloalkyl, haloalkenyl, haloalkynyl, halocycloalkyl,halocycloalkenyl, alkoxy, alkenyloxy, alkynyloxy, haloalkoxy,haloalkenyloxy, haloalkynyloxy, cycloalkoxy, cycloalkenyloxy,halocycloalkoxy, halocycloalkenyloxy, alkylthio, haloalkylthio,cycloalkylthio, halocycloalkylthio, alkylsulfinyl, alkenylsulfinyl,alkynyl-sulfinyl, haloalkylsulfinyl, haloalkenylsulfinyl,haloalkynylsulfinyl, alkylsulfonyl, alkenylsulfonyl, alkynylsulfonyl,haloalkyl-sulfonyl, haloalkenylsulfonyl, haloalkynylsulfonyl,alkylamino, alkenylamino, alkynylamino, di(alkyl)amino,di(alkenyl)-amino, di(alkynyl)amino, or trialkylsilyl.

The term “alkylaryl,” as used herein, refers to an aryl group that isbonded to a parent compound through a diradical alkylene bridge,(—CH₂-)_(n), where n is 1-12 and where “aryl” is as defined above.

The term “alkylcycloalkyl,” as used herein, refers to a cycloalkyl groupthat is bonded to a parent compound through a diradical alkylene bridge,(—CH₂-)_(n), where n is 1-12 and where “cycloalkyl” is as defined above.The term “cycloalkylalkyl,” as used herein, refers to a cycloalkylgroup, as defined above, which is substituted by an alkyl group, asdefined above.

The term “heteroalkyl,” as used herein, refers to an alkyl group, asdescribed above, which includes one or more heteroatoms (e.g., from oneto four heteroatoms) within the carbon backbone. In some cases, theheteroatom(s) incorporated into the carbon backbone are oxygen,nitrogen, sulfur, or combinations thereof. The terms “heteroalkenyl” and“heteroalkynyl,” as used herein, likewise refer to alkenyl and alkynylgroups respectively which include one or more heteroatoms (e.g., fromone to four heteroatoms, such as oxygen, nitrogen, sulfur, orcombinations thereof) within their carbon backbone.

The term “heteroaryl,” as used herein, refers to a monovalent aromaticgroup of from 1 to 15 carbon atoms (e.g., from 1 to 10 carbon atoms,from 2 to 8 carbon atoms, from 3 to 6 carbon atoms, or from 4 to 6carbon atoms) having one or more heteroatoms within the ring. Theheteroaryl group can include from 1 to 4 heteroatoms, from 1 to 3heteroatoms, or from 1 to 2 heteroatoms. In some cases, theheteroatom(s) incorporated into the ring are oxygen, nitrogen, sulfur,or combinations thereof. When present, the nitrogen and sulfurheteroatoms may optionally be oxidized. Heteroaryl groups can have asingle ring (e.g., pyridyl or furyl) or multiple condensed ringsprovided that the point of attachment is through a heteroaryl ring atom.Preferred heteroaryls include pyridyl, piridazinyl, pyrimidinyl,pyrazinyl, triazinyl, pyrrolyl, indolyl, quinolinyl, isoquinolinyl,quinazolinyl, quinoxalinnyl, furanyl, thiophenyl, furyl, pyrrolyl,imidazolyl, oxazolyl, isoxazolyl, isothiazolyl, pyrazolyl benzofuranyl,and benzothiophenyl. Heteroaryl rings may be unsubstituted orsubstituted by one or more moieties as described for aryl above.

The term “alkylheteroaryl,” as used herein, refers to a heteroaryl groupthat is bonded to a parent compound through a diradical alkylene bridge,(—CH₂-)_(n), where n is 1-12 and where “heteroaryl” is as defined above.

The terms “cycloheteroalkyl,” “heterocyclyl,” “heterocyclic,” and“heterocyclo” are used herein interchangeably, and refer to fullysaturated or unsaturated, cyclic groups, for example, 3 to 7 memberedmonocyclic or 4 to 7 membered monocyclic; 7 to 11 membered bicyclic, or10 to 15 membered tricyclic ring systems, having one or more heteroatomswithin the ring. The heterocyclyl group can include from 1 to 4heteroatoms, from 1 to 3 heteroatoms, or from 1 to 2 heteroatoms. Insome cases, the heteroatom(s) incorporated into the ring are oxygen,nitrogen, sulfur, or combinations thereof. When present, the nitrogenand sulfur heteroatoms may optionally be oxidized, and the nitrogenheteroatoms may optionally be quaternized. The heterocyclyl group may beattached at any heteroatom or carbon atom of the ring or ring system andmay be unsubstituted or substituted by one or more moieties as describedfor aryl groups above.

Exemplary monocyclic heterocyclic groups include, but are not limitedto, pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanyl, pyrazolinyl,imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl,isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl,isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl,oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl,4-piperidonyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl,tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinylsulfoxide, thiamorpholinyl sulfone, 1,3-dioxolane andtetrahydro-1,1-dioxothienyl, triazolyl, triazinyl, and the like.

Exemplary bicyclic heterocyclic groups include, but are not limited to,indolyl, benzothiazolyl, benzoxazolyl, benzodioxolyl, benzothienyl,quinuclidinyl, quinolinyl, tetra-hydroisoquinolinyl, isoquinolinyl,benzimidazolyl, benzopyranyl, indolizinyl, benzofuryl, chromonyl,coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl,pyrrolopyridyl, furopyridinyl (such as furo[2,3-c]pyridinyl,furo[3,2-b]pyridinyl] or furo[2,3-b]pyridinyl), dihydroisoindolyl,dihydroquinazolinyl (such as 3,4-dihydro-4-oxo-quinazolinyl),tetrahydroquinolinyl and the like.

Exemplary tricyclic heterocyclic groups include carbazolyl, benzidolyl,phenanthrolinyl, acridinyl, phenanthridinyl, xanthenyl, and the like.

The term “alkylheterocyclyl” and “alkylcycloheteroalkyl” are used hereininterchangeably, and refer to a heterocyclyl group that is bonded to aparent compound through a diradical alkylene bridge, (—CH₂-)_(n), wheren is 1-12 and where “heterocyclyl” is as defined above. The term“heterocyclylalkyl,” as used herein, refers to a heterocyclyl group, asdefined above, which is substituted by an alkyl group, as defined above.

The term “halogen,” as used herein, refers to the atoms fluorine,chlorine, bromine and iodine. The prefix halo- (e.g., as illustrated bythe term haloalkyl) refers to all degrees of halogen substitution, froma single substitution to a perhalo substitution (e.g., as illustratedwith methyl as chloromethyl (—CH₂Cl), dichloromethyl (—CHCl₂),trichloromethyl (—CCl₃)).

As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, and aromatic and nonaromaticsubstituents of organic compounds. Illustrative substituents include,for example, those described below. The permissible substituents can beone or more and the same or different for appropriate organic compounds.For purposes of this disclosure, the heteroatoms, such as nitrogen, canhave hydrogen substituents and/or any permissible substituents oforganic compounds described herein which satisfy the valences of theheteroatoms. This disclosure is not intended to be limited in any mannerby the permissible substituents of organic compounds. Also, the terms“substitution” or “substituted with” include the implicit proviso thatsuch substitution is in accordance with permitted valence of thesubstituted atom and the substituent, and that the substitution resultsin a stable compound, e.g., a compound that does not spontaneouslyundergo transformation such as by rearrangement, cyclization,elimination, etc.

Stereoisomers and Polymorphic Forms

Unless stated to the contrary, a formula with chemical bonds shown onlyas solid lines and not as wedges or dashed lines contemplates eachpossible isomer, e.g., each enantiomer, diastereomer, and meso compound,and a mixture of isomers, such as a racemic or scalemic mixture.

The compounds described herein can exist and be isolated as opticallyactive and racemic forms. The compounds can have one or more chiralcenters, including at a sulfur atom, and thus exist as one or morestereoisomers. Where compounds include n chiral centers, the compoundscan comprise up to 2^(n) optical isomers. Such stereoisomer-containingcompounds can exist as a single enantiomer, a mixture of enantiomers, amixture of diastereomers, or a racemic mixture. The optically activeforms can be prepared by, for example, resolution of the racemic formsby selective crystallization techniques, by synthesis from opticallyactive precursors, by chiral synthesis, by chromatographic separationusing a chiral stationary phase or by enzymatic resolution.

The compounds can also be present in different solid forms, includingdifferent crystalline forms (i.e., different crystalline polymorphs ofthe compounds) or as an amorphous solid. In addition, the compounds canexist as hydrates or solvates, in which a certain stoichiometric amountof water or a solvent is associated with the molecule in the crystallineform. In some embodiments, the compositions described herein can includeup to 15% (w/w), up to 20% (w/w), or up to 30% (w/w) of a particularsolid form of the compounds described herein, based on the total weightof the composition.

Pharmaceutically Acceptable Salts

The compounds described herein can also be provided as pharmaceuticallyacceptable salts (e.g., acid or base salts) where applicable, of thecompounds described herein. Pharmaceutically acceptable salts are knownin the art. See, for example, Remington's Pharmaceutical Sciences, 20thed., Lippincott Williams & Wilkins, Baltimore, Md., 2000, p. 704.

The term “acid salt” contemplates salts of the compounds with allpharmaceutically acceptable inorganic or organic acids. Inorganic acidsinclude mineral acids such as hydrohalic acids such as hydrobromic acidand hydrochloric acid, sulfuric acid, phosphoric acids and nitric acid.Organic acids include all pharmaceutically acceptable aliphatic,alicyclic and aromatic carboxylic acids, dicarboxylic acids,tricarboxylic acids and fatty acids. In one embodiment of the acids, theacids are straight chain or branched, saturated or unsaturated C₁-C₂₀aliphatic carboxylic acids, which are optionally substituted by halogenor by hydroxyl groups, or C₆-C₁₂ aromatic carboxylic acids. Examples ofsuch acids are carbonic acid, formic acid, acetic acid, propionic acid,isopropionic acid, valeric acid, α-hydroxy acids such as glycolic acidand lactic acid, chloroacetic acid, benzoic acid, methane sulfonic acid,and salicylic acid. Examples of dicarboxylic acids include oxalic acid,malic acid, succinic acid, tartaric acid, fumaric acid, and maleic acid.An example of a tricarboxylic acid is citric acid. Fatty acids includeall pharmaceutically acceptable saturated or unsaturated aliphatic oraromatic carboxylic acids having 4 to 24 carbon atoms. Examples includebutyric acid, isobutyric acid, sec-butyric acid, lauric acid, palmiticacid, stearic acid, oleic acid, linoleic acid, linolenic acid, andphenylsteric acid. Other acids include gluconic acid, glycoheptonic acidand lactobionic acid.

The term “base salt” contemplates salts of the compounds with allpharmaceutically acceptable inorganic or organic bases, includinghydroxides, carbonates or bicarbonates of alkali metal or alkaline earthmetals. Salts formed with such bases include, for example, the alkalimetal and alkaline earth metal salts, including, but not limited to, asthe lithium, sodium, potassium, magnesium or calcium salts. Salts formedwith organic bases include the common hydrocarbon and heterocyclic aminesalts, which include, for example, ammonium salts (NH₄ ⁺), alkylammoniumsalts, and dialkylammonium salts, as well as salts of cyclic amines suchas the morpholine and piperidine salts.

Prodrugs

The compounds described herein can also be provided as pharmaceuticallyacceptable prodrugs. Prodrugs of are compounds that, when metabolized invivo, undergo conversion to compounds described herein having thedesired pharmacological activity. Prodrugs can be prepared by replacingappropriate functionalities present in the compounds described hereinwith “pro-moieties” as described, for example, in H. Bundgaar, Design ofProdrugs (1985). Examples of prodrugs include ester, ether or amidederivatives of the compounds described herein, as well as theirpharmaceutically acceptable salts. For further discussions of prodrugs,see, for example, T. Higuchi and V. Stella “Pro-drugs as Novel DeliverySystems,” ACS Symposium Series 14 (1975) and E. B. Roche ed.,Bioreversible Carriers in Drug Design (1987).

Compounds

Provided herein are compounds that can exhibit activity as biofilmmodulating agents (e.g., activity as biofilm inhibitors and/or activityas biofilm dispersal agents). The compounds can exhibit potent activityagainst Gram positive biofilms. The compounds can also exhibit activityagainst Gram negative biofilms. In some cases, the compounds can exhibitboth biofilm modulation properties and antimicrobial activity.

For example, provided herein are compounds defined by Formula I

or a pharmaceutically acceptable salt or prodrug thereof, wherein X ischosen from O and S; R¹, R², R³, R⁴, and R⁵ are each independentlychosen from hydrogen, halogen, hydroxyl, —CN, —NO₂, amino, alkylamino,dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy,haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl,haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl,haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁶ and R⁷ are each independently chosen fromhydrogen, hydroxy, halogen, —CN, —NO₂, amino, alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl, haloalkylsulfinyl,alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl,alkoxycarbonyl, haloalkoxycarbonyl, alkylaminocarbonyl,heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁸ is chosen from alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, alkylcycloheteroalkyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl, each optionallysubstituted with one or more substituents individually chosen from R¹¹;R⁹ is chosen from hydrogen, hydroxy, halogen, —CN, —NO₂, amino,alkylamino, dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio,alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, aryl,heteroaryl, cycloalkyl, cycloheteroalkyl, alkylaryl, alkylheteroaryl,alkylcycloalkyl, alkylcycloheteroalkyl, alkylsulfinyl,haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl,haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl, each optionally substituted with one or moresubstituents individually chosen from R¹¹; R¹⁰ is chosen from hydrogen,alkyl, haloalkyl, alkylthio, haloalkylthio, alkenyl, haloalkenyl,alkynyl, haloalkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, and alkylcycloheteroalkyl,each optionally substituted with one or more substituents individuallychosen from R¹¹; and R¹¹ is chosen from hydroxy, halogen, —CN, —NO₂,amino, alkylamino, dialkylamino, alkyl, haloalkyl; alkylthio;haloalkylthio; alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl. In someembodiments, the compound is not one of the following:

In some embodiments, X can be O.

In some embodiments, at least one of R¹, R², R³, R⁴, and R⁵ is nothydrogen. In some cases, R¹, R², R⁴, and R⁵ are hydrogen. In certaincases, R¹, R², R⁴, and R⁵ are hydrogen, and R⁴ is not hydrogen (e.g.,the phenyl ring of the benzylidene moiety is para-substituted).

In some embodiment, R³ can be an electron withdrawing group. Forexample, R³ can be chosen from halogen, —CN, —NO₂, haloalkyl,alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl, andhaloalkoxycarbonyl. In some embodiments, R³ can be a haloalkyl group. Incertain embodiments, R³ can be a perfluoroalkyl group (e.g., a —CF₃group).

In some embodiments, at least one of R⁶ and R⁷ is not hydrogen. Incertain embodiments, both R⁶ and R⁷ are not hydrogen. In some cases, atleast one of R⁶ and R⁷ can be halogen (e.g., at least one of R⁶ and R⁷can be —Cl and/or at least one of R⁶ and R⁷ can be —F). In some cases,both R⁶ and R⁷ can be halogen (e.g., both R⁶ and R⁷ can be —C₁, or bothR⁶ and R⁷ can be —F).

In some examples, R⁸ can comprise a substituent having from 5 to 12carbon atoms. In certain examples, R⁸ can be chosen from alkyl, aryl,heteroaryl, alkylaryl, and alkylheteroaryl. In some embodiments, R⁸ cancomprise a C₁-C₈ alkyl group (e.g., a C₁-C₄ alkyl group, or a C₅-C₈alkyl group). In other embodiments, R⁸ can comprise a C₆-C₁₀ alkylarylgroup (e.g., a substituted or unsubstituted benzyl group).

In some embodiments, R⁹ can be hydrogen. In other embodiments, R⁹ cancomprise an alkyl group (e.g., a C₁-C₄ alkyl group) or an aryl group(e.g., a phenyl group).

In some embodiments, R¹⁰ can be hydrogen. In other embodiments, R¹⁰ canbe an alkyl group, an aryl group, or an alkylaryl group. In certainembodiments, R10 can comprise an alkyl group (e.g., a C₁-C₄ alkyl group)or an aryl group (e.g., a phenyl group).

In some cases, the compound can be a compound defined by Formula II

or a pharmaceutically acceptable salt or prodrug thereof, wherein R³comprises an electron withdrawing group; R⁶ and R⁷ are eachindependently chosen from hydrogen and halogen, with the proviso that atleast one of R⁶ and R⁷ is halogen; R⁸ is chosen from alkyl, haloalkyl,alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, aryl,heteroaryl, cycloalkyl, cycloheteroalkyl, alkylaryl, alkylheteroaryl,alkylcycloalkyl, alkylcycloheteroalkyl, each optionally substituted withone or more substituents individually chosen from R¹¹; and R¹¹ is chosenfrom hydroxy, halogen, —CN, —NO₂, amino, alkylamino, dialkylamino,alkyl, haloalkyl; alkylthio; haloalkylthio; alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl, haloalkylsulfinyl,alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl,alkoxycarbonyl, haloalkoxycarbonyl, alkylaminocarbonyl,heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl. In some embodiments, the compound is not oneof the following:

In some embodiments, R³ can be chosen from halogen, —CN, —NO₂,haloalkyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl, andhaloalkoxycarbonyl. In some embodiments, R³ can be a haloalkyl group. Incertain embodiments, R³ can be a perfluoroalkyl group (e.g., a —CF₃group).

In some embodiments, the compound can be one of the following:

In some embodiments, the compound can be one of the following:

Compositions

Also provided are compositions that include one or more of the compoundsdescribed herein. In some embodiments, biofilm preventing, removing orinhibiting compositions are provided, comprising a carrier and aneffective amount of a compound described herein.

In some embodiments, the carrier can be a pharmaceutically acceptablecarrier. A “pharmaceutically acceptable carrier” as used herein refersto a carrier that, when combined with a compound described herein,facilitates the application or administration of that compound describedherein for its intended purpose (e.g., to prevent or inhibit biofilmformation, or remove an existing biofilm). The compound described hereinmay be formulated for administration in a pharmaceutically acceptablecarrier in accordance with known techniques. See, e.g., Remington, TheScience And Practice of Pharmacy (9th Ed. 1995). The pharmaceuticallyacceptable carrier can, of course, also be acceptable in the sense ofbeing compatible with any other ingredients in the composition.

The carrier may be a solid or a liquid, or both, and is preferablyformulated with the a compound described herein as a unit-dosecomposition, for example, a tablet, which may contain from 0.01 or 0.5%to 95% or 99% by weight of the a compound described herein. One or morea compounds described herein can be included in the compositions, whichmay be prepared by any of the well-known techniques of pharmacycomprising admixing the components, optionally including one or moreaccessory ingredients.

In general, compositions may be prepared by uniformly and intimatelyadmixing the a compound described herein with a liquid or finely dividedsolid carrier, or both, and then, if necessary, shaping the resultingmixture. For example, a tablet may be prepared by compressing or moldinga powder or granules containing the a compound described herein,optionally with one or more accessory ingredients. Compressed tabletsmay be prepared by compressing, in a suitable machine, the compound in afree-flowing form, such as a powder or granules optionally mixed with abinder, lubricant, inert diluent, and/or surface active/dispersingagent(s). Molded tablets may be made by molding, in a suitable machine,the powdered compound moistened with an inert liquid binder.

Compositions can be formulated to be suitable for oral, rectal, topical,buccal (e.g., sub-lingual), vaginal, parenteral (e.g., subcutaneous,intramuscular, intradermal, or intravenous), topical (i.e., both skinand mucosal surfaces, including airway surfaces) or transdermaladministration, although the most suitable route in any given case willdepend on the nature and severity of the condition being treated and onthe nature of the particular compound that is being used.

Compositions suitable for oral administration may be presented indiscrete units, such as capsules, cachets, lozenges, or tablets, eachcontaining a predetermined amount of the compound; as a powder orgranules; as a solution or a suspension in an aqueous or non-aqueousliquid; or as an oil-in-water or water-in-oil emulsion. Suchcompositions may be prepared by any suitable method of pharmacy, whichincludes the step of bringing into association the compound and asuitable carrier (which may contain one or more accessory ingredients asnoted above).

Compositions suitable for buccal (sub-lingual) administration includelozenges comprising the compound in a flavored base, usually sucrose andacacia or tragacanth; and pastilles comprising the compound in an inertbase such as gelatin and glycerin or sucrose and acacia.

Compositions suitable for parenteral administration comprise sterileaqueous and non-aqueous injection solutions of the compound, whichpreparations are preferably isotonic with the blood of the intendedrecipient. These preparations may contain anti-oxidants, buffers,bacteriostats and solutes that render the composition isotonic with theblood of the intended recipient. Aqueous and non-aqueous sterilesuspensions may include suspending agents and thickening agents. Thecompositions may be presented in unit/dose or multi-dose containers, forexample sealed ampoules and vials, and may be stored in a freeze-dried(lyophilized) condition requiring only the addition of the sterileliquid carrier, for example, saline or water-for-injection immediatelyprior to use. Extemporaneous injection solutions and suspensions may beprepared from sterile powders, granules and tablets of the kindpreviously described.

For example, the composition can be an injectable, stable, sterilecomposition comprising a compound described herein in a unit dosage formin a sealed container. The compostion can be provided in the form of alyophilizate that can be reconstituted with a suitable pharmaceuticallyacceptable carrier to form a liquid composition suitable for injectionthereof into a subject. The unit dosage form can comprise from about 10mg to about 10 grams of the compound. When the compound or salt issubstantially water-insoluble, a sufficient amount of emulsifying agentthat is physiologically acceptable may be employed in sufficientquantity to emulsify the compound or salt in an aqueous carrier. Onesuch useful emulsifying agent is phosphatidyl choline.

Compositions suitable for rectal administration can be presented as unitdose suppositories. These may be prepared by mixing the active compoundwith one or more conventional solid carriers, for example, cocoa butter,and then shaping the resulting mixture.

Compositions suitable for topical application to the skin can take theform of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil.Carriers that may be used include petroleum jelly, lanoline,polyethylene glycols, alcohols, transdermal enhancers, and combinationsof two or more thereof.

Compositions suitable for transdermal administration can be presented asdiscrete patches adapted to remain in intimate contact with theepidermis of the recipient for a prolonged period of time. Compositionssuitable for transdermal administration may also be delivered byiontophoresis and typically take the form of an optionally bufferedaqueous solution of the active compound.

In some embodiments, the compositions described herein can furtherinclude one or more additional active agents, such as a biocide. A“biocide” as used herein refers to a substance with the ability to killor to inhibit the growth of microorganisms (e.g., bacteria, fungalcells, protozoa, etc.), which is not compound described in the compoundssection above. Common biocides include oxidizing and non-oxidizingchemicals.

In some embodiments, the compositions described herein can furtherinclude one or more antibiotics. An “antibiotic” as used herein is atype of “biocide.” Common antibiotics include aminoglycosides,carbacephems (e.g., loracarbef), carbapenems, cephalosporins,glycopeptides (e.g., teicoplanin and vancomycin), macrolides,monobactams (e.g., aztreonam) penicillins, polypeptides (e.g.,bacitracin, colistin, polymyxin B), quinolones, sulfonamides,tetracyclines, etc. Antibiotics treat infections by either killing orpreventing the growth of microorganisms. Many act to inhibit cell wallsynthesis or other vital protein synthesis of the microorganisms.

Aminoglycosides are commonly used to treat infections caused byGram-negative bacteria such as Escherichia coli and Klebsiella,particularly Pseudomonas aeroginosa. Examples of aminoglycosidesinclude, but are not limited to amikacin, gentamicin, kanamycin,neomycin, netilmicin, streptomycin, tobramycin, and paromomycin.

Carbapenems are broad-specrum antibiotics, and include, but are notlimited to, ertapenem, doripenem, imipenem/cilstatin, and meropenem.

Cephalosporins include, but are not limited to, cefadroxil, cefazolin,cefalotin (cefalothin), cefalexin, cefaclor, cefamandole, cefoxitin,cefprozil, loracarbef, cefuroxime, cefixime, cefdinir, cefditoren,cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten,ceftizoxime, ceftriaxone, cefepime, cefpirome, and ceftobiprole.

Macrolides include, but are not limited to, azithromycin,clarithromycin, dirithromycin, erythromycin, roxithromycin,troleandomycin, telithromycin and spectinomycin.

Penicillins include, but are not limited to, amoxicillin, ampicillin,azlocillin, bacampicillin, carbenicillin, cloxacillin, dicloxacillin,flucloxacillin, mezlocillin, meticillin, nafcillin, oxacillin,penicillin, piperacillin and ticarcillin.

Quinolones include, but are not limited to, ciprofloxacin, enoxacin,gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin,norfloxacin, ofloxacin and trovafloxacin.

Sulfonamides include, but are not limited to, mafenide, prontosil,sulfacetamide, sulfamethizole, sulfanilamide, sulfasalazine,sulfisoxazole, trimethoprim, and co-trimoxazole(trimethoprim-sulfamethoxazole).

Tetracyclines include, but are not limited to, demeclocycline,doxycycline, minocycline, oxytetracycline and tetracycline.

Other antibiotics include arsphenamine, chloramphenicol, clindamycin,lincomycin, ethambutol, fosfomycin, fusidic acid, furazolidone,isoniazid, linezolid, metronidazole, mupirocin, nitrofurantoin,platensimycin, pyrazinamide, quinupristin/dalfopristin, rifampin(rifampicin), tinidazole, etc.

In some embodiments, the composition can be a dentifrice compositioncomprising one or more of the compounds described herein. A “dentifrice”is a substance that is used to clean the teeth. It may be in the formof, e.g., a paste or powder. Commonly known dentifrices includetoothpaste, mouthwash, chewing gum, dental floss, and dental cream.Other examples of dentifrices include toothpowder, mouth detergent,troches, dental or gingival massage cream, dental strips, dental gels,and gargle tablets. Examples of dentifrice compositions comprisingtoothpaste and mouthwash are found in U.S. Pat. No. 6,861,048 (Yu etal.); U.S. Pat. No. 6,231,836 (Takhtalian et al.); and U.S. Pat. No.6,331,291 (Glace et al.); each of which are incorporated by referenceherein in their entirety.

Coating compositions are also provided. A “coating” as used herein isgenerally known. Any of a variety of organic and aqueous coatingcompositions, with or without pigments, may be modified to contain oneor more compounds described herein. Examples of suitable coatingcompositions include, for example, the coating compositions described inU.S. Pat. Nos. 7,109,262, 6,964,989, 6,835,459, 6,677,035, 6,528,580,and 6,235,812, each incorporated by reference herein in their entirety.

In some examples, coating compositions can comprise (in addition to oneor more compounds described herein) a film-forming resin, an aqueous ororganic solvent that disperses the resin; and, optionally, at least onepigment. Other ingredients such as colorants, secondary pigments,stabilizers and the like can be included if desired. The one or morebiofilm modulating compounds described herein may be dissolved ordispersed in the solvent and/or resin, so that the compound(s) aredispersed or distributed on the substrate an article coated by thecoating composition. The resin may comprise, for example, a polymericmaterial. A polymeric material is a material that is comprised of largemolecules made from associated smaller repeating structural units, oftencovalently linked. Common examples of polymeric materials areunsaturated polyester resins, and epoxy resins.

Any suitable article can be coated, in whole or in part, with thecoating compositions described herein. Suitable articles include, butare not limited to, automobiles and airplanes (including substrates suchas wing and propeller surfaces for aerodynamic testing), vessel hulls(including interior and exterior surfaces thereof), pressure vessels(including interior and exterior surfaces thereof), medical devices(e.g., implants), windmills, etc. Coating of the article with thecomposition can be carried out by any suitable means, such as bybrushing, spraying, electrostatic deposition, dip coating, doctorblading, etc.

Devices

Also provided are medical devices that comprise a medical devicesubstrate and an effective amount of a compound described herein eithercoating the substrate, or incorporated into the substrate. The effectiveamount of the compound can be an effective amount to prevent or inhibitgrowth of a biofilm on the medical device substrate.

“Medical device” as used herein refers to an object that is inserted orimplanted in a subject or applied to a surface of a subject. Commonexamples of medical devices include stents, fasteners, ports, catheters,scaffolds and grafts. A “medical device substrate” can be made of avariety of biocompatible materials, including, but not limited to,metals, ceramics, polymers, gels, and fluids not normally found withinthe human body. Examples of polymers useful in fabricating medicaldevices include such polymers as silicones, rubbers, latex, plastics,polyanhydrides, polyesters, polyorthoesters, polyamides,polyacrylonitrile, polyurethanes, polyethylene, polytetrafluoroethylene,polyethylenetetraphthalate, etc. Medical devices can also be fabricatedusing naturally-occurring materials or treated with naturally-occurringmaterials. Medical devices can include any combination of artificialmaterials, e.g., combinations selected because of the particularcharacteristics of the components. Medical devices can be intended forshort-term or long-term residence where they are positioned. A hipimplant is intended for several decades of use, for example. Bycontrast, a tissue expander may only be needed for a few months, and isremoved thereafter.

Some examples of medical devices are found in U.S. Pat. No. 7,081,133(Chinn et al.); U.S. Pat. No. 6,562,295 (Neuberger); and U.S. Pat. No.6,387,363 (Gruskin); each incorporated by reference herein in theirentirety.

Methods of Use

Methods of controlling biofilm formation on a substrate are disclosed,comprising the step of administering a compound described herein to asubstrate in an amount effective to inhibit biofilm formation.

A “substrate” as used herein is a base on which an organism, such asthose commonly found in biofilms, may live. The term “substrate,” asused herein, refers to any substrate, whether in an industrial or amedical setting, that provides or can provide an interface between anobject and a fluid, permitting at least intermittent contact between theobject and the fluid. A substrate, as understood herein, furtherprovides a plane whose mechanical structure, without further treatment,is compatible with the adherence of microorganisms. Substratescompatible with biofilm formation may be natural or synthetic, and maybe smooth or irregular. Fluids contacting the substrates can be stagnantor flowing, and can flow intermittently or continuously, with laminar orturbulent or mixed flows. A substrate upon which a biofilm forms can bedry at times with sporadic fluid contact, or can have any degree offluid exposure including total immersion. Fluid contact with thesubstrate can take place via aerosols or other means for air-borne fluidtransmission.

Biofilm formation with health implications can involve those substratesin all health-related environments, including substrates found inmedical environments and those substrates in industrial or residentialenvironments that are involved in those functions essential to humanwell being, for example, nutrition, sanitation and the prevention ofdisease. Substrates found in medical environments include the inner andouter aspects of various instruments and devices, whether disposable orintended for repeated uses. Examples include the entire spectrum ofarticles adapted for medical use, including scalpels, needles, scissorsand other devices used in invasive surgical, therapeutic or diagnosticprocedures; implantable medical devices, including artificial bloodvessels, catheters and other devices for the removal or delivery offluids to patients, artificial hearts, artificial kidneys, orthopedicpins, plates and implants; catheters and other tubes (includingurological and biliary tubes, endotracheal tubes, peripherablyinsertable central venous catheters, dialysis catheters, long termtunneled central venous catheters, peripheral venous catheters, shortterm central venous catheters, arterial catheters, pulmonary catheters,Swan-Ganz catheters, urinary catheters, peritoneal catheters), urinarydevices (including long term urinary devices, tissue bonding urinarydevices, artificial urinary sphincters, urinary dilators), shunts(including ventricular or arterio-venous shunts); prostheses (includingbreast implants, penile prostheses, vascular grafting prostheses, heartvalves, artificial joints, artificial larynxes, otological implants),vascular catheter ports, wound drain tubes, hydrocephalus shunts,pacemakers and implantable defibrillators, and the like. Other exampleswill be readily apparent to practitioners in these arts. Substratesfound in the medical environment also include the inner and outeraspects of pieces of medical equipment, medical gear worn or carried bypersonnel in the health care setting. Such substrates can includecounter tops and fixtures in areas used for medical procedures or forpreparing medical apparatus, tubes and canisters used in respiratorytreatments, including the administration of oxygen, of solubilized drugsin nebulizers and of anesthetic agents. Also included are thosesubstrates intended as biological barriers to infectious organisms inmedical settings, such as gloves, aprons and faceshields. Commonly usedmaterials for biological barriers may be latex-based or non-latex based.Vinyl is commonly used as a material for non-latex surgical gloves.Other such substrates can include handles and cables for medical ordental equipment not intended to be sterile. Additionally, suchsubstrates can include those non-sterile external substrates of tubesand other apparatus found in areas where blood or body fluids or otherhazardous biomaterials are commonly encountered.

Substrates in contact with liquids are particularly prone to biofilmformation. As an example, those reservoirs and tubes used for deliveringhumidified oxygen to patients can bear biofilms inhabited by infectiousagents. Dental unit waterlines similarly can bear biofilms on theirsubstrates, providing a reservoir for continuing contamination of thesystem of flowing an aerosolized water used in dentistry. Sprays,aerosols and nebulizers are highly effective in disseminating biofilmfragments to a potential host or to another environmental site. It isespecially important to health to prevent biofilm formation on thosesubstrates from where biofilm fragments can be carried away by sprays,aerosols or nebulizers contacting the substrate.

Other substrates related to health include the inner and outer aspectsof those articles involved in water purification, water storage andwater delivery, and articles involved in food processing. Substratesrelated to health can also include the inner and outer aspects of thosehousehold articles involved in providing for nutrition, sanitation ordisease prevention. Examples can include food processing equipment forhome use, materials for infant care, tampons and toilet bowls.“Substrate” as used herein also refers to a living substrate, such asthe inner ear of a patent.

Substrates can be smooth or porous, soft or hard. Substrates can includea drainpipe, glaze ceramic, porcelain, glass, metal, wood, chrome,plastic, vinyl, Formica® brand laminate, or any other material that mayregularly come in contact with an aqueous solution in which biofilms mayform and grow. The substrate can be a substrate commonly found onhousehold items such as shower curtains or liners, upholstery, laundry,and carpeting.

A substrate on which biofilm preventing, removing or inhibiting isimportant is that of a ship hull. Biofilms, such as those of Halomonaspacifica, promote the corrosion of the hull of ships and also increasethe roughness of the hull, increasing the drag on the ship and therebyincreasing fuel costs. The biofilm can also promote the attachment oflarger living structures such as barnacles on the ship hull. Fuel canaccount for half of the cost of marine shipping, and the loss in fuelefficiency due to biofilm formation is substantial.

Substrates on which biofilms can adhere include those of livingorganisms, as in the case of humans with chronic infections caused bybiofilms, as discussed above. Biofilms can also form on the substratesof food contact surfaces, such as those used for processing seafood, andalso on food products themselves. Examples of seafood products that mayhave biofilm contamination include oysters. Human infections caused bythe ingestion of raw oysters has been linked to Vibrio vulnificusbacterium. Vibrio bacteria attach to algae and plankton in the water andtransfer to the oysters and fish that feed on these organisms.

Other examples of substrates or devices on which biofilms can adhere canbe found in U.S. Pat. Nos. 5,814,668 and 7,087,661; and U.S. Pat.Application Publication Nos. 2006/0228384 and 2006/0018945, each ofwhich is incorporated herein by reference in its entirety.

In some embodiments, methods of enhancing the effects of a biocide aredisclosed, comprising the step of administering a compound describedherein in combination with a biocide, the active compound beingadministered in an amount effective to enhance the effects of thebiocide.

“Administering” or “administration of” a compound described hereinand/or biocide as used herein in inclusive of contacting, applying, etc.(e.g., contacting with an aqueous solution, contacting with a surface(e.g., a hospital surface such as a table, instrumentation, etc.)), inaddition to providing to a subject (for example, to a human subject inneed of treatment for a microbial infection).

“Enhancing” the effects of a biocide by administering a compounddescribed herein in combination with the biocide refers to increasingthe effectiveness of the biocide, such that the microorganism killingand/or growth inhibition is higher at a certain concentration of thebiocide administered in combination with the active compound thanwithout. In some embodiments, a bacteria or other microorganism is“sensitized” to the effects of a biocide, such that the bacteria orother microorganism that was resistant to the biocide prior toadministering the compound described herein (e.g., little to none, orless than 20, 10, 5 or 1% are killed upon application) is renderedvulnerable to that biocide upon or after administering the compound(e.g., greater than 20, 30, 40, 50, 60, 70, 80, 90, or 95% or more arekilled).

As used herein, the administration of two or more compounds (inclusiveof the compounds described herein and biocides) “in combination” meansthat the two compounds are administered closely enough in time that theadministration of or presence of one alters the biological effects ofthe other. The two compounds may be administered simultaneously(concurrently) or sequentially.

Simultaneous administration of the compounds may be carried out bymixing the compounds prior to administration, or by administering thecompounds at the same point in time but at different anatomic sites orusing different routes of administration, or administered at timessufficiently close that the results observed are indistinguishable fromthose achieved when the compounds are administered at the same point intime.

Sequential administration of the compounds may be carried out byadministering, e.g., an active compound at some point in time prior toadministration of a biocide, such that the prior administration ofactive compound enhances the effects of the biocide (e.g., percentage ofmicroorganisms killed and/or slowing the growth of microorganisms). Insome embodiments, an active compound is administered at some point intime prior to the initial administration of a biocide. Alternatively,the biocide may be administered at some point in time prior to theadministration of an active compound, and optionally, administered againat some point in time after the administration of an active compound.

Also provided herein are methods for controlling biofilm formation on asubstrate. Methods for controlling biofilm formation on a substrate cancomprise contacting the substrate with a compound described herein in anamount effective to inhibit biofilm formation.

The biofilm can comprise Gram-positive bacteria or Gram-negativebacteria. In some embodiments, the biofilm can comprise Gram-positivebacteria. Examples of Gram-positive bacteria affected by compoundsdescribed herein include, but are not limited to, bacteria of the generaListeria, Staphylococcus, Streptococcus, Bacillus, Corynebacterium,Peptostreptococcus, and Clostridium. For example, the bacteria caninclude Listeria monocytogenes, Staphylococcus aureus, Streptococcuspyogenes, Streptococcus pneumoniae, Bacillus cereus, Bacillus anthracis,Clostridium botulinum, Clostridium perfringens, Clostridium difficile,Clostridium tetani, Corynebacterium diphtheriae, Corynebacteruimulcerans, and Peptostreptococcus anaerobius. Other examples ofGram-positive bacteria include, for example, bacteria of the generaActinomyces, Propionibacterium, Nocardia and Streptomyces.

In some embodiments, the biofilm can comprise Gram-negative bacteria.Examples of Gram-positive bacteria affected by compounds describedherein include, but are not limited to, bacteria of the generaEscherichia, Salmonella, Vibrio, Helicobacter, Pseudomonas, Bordetella,Vibrio, Haemophilus, Halomonas, and Acinetobacter. For example, thebacteria can include Pseudomonas aeuroginosa, Bordetella pertussis,Vibrio vulnificus, Haemophilus influenzae, Halomonas pacifica, andAcinetobacter baumannii. Other examples of Gram-negative bacteriainclude, for example, bacteria of the genera Klebsiella, Proteus,Neisseria, Helicobacter; Brucella, Legionella, Campylobacter,Francisella, Pasteurella, Yersinia, Bartonella, Bacteroides,Streptobacillus, Spirillum, Moraxella and Shigella.

Also provided are methods for treating chronic bacterial infections in asubject in need thereof. These methods can comprise administering acompound described herein to a subject in an amount effective toinhibit, reduce, or remove a biofilm component of said chronic bacterialinfection. “Treating” as used herein refers to any type of activity thatimparts a benefit to a patient afflicted with a disease, includingimprovement in the condition of the patient (e.g., in one or moresymptoms), delay in the progression of the disease, delay in onset ofthe disease, etc.

A “chronic bacterial infection” is a bacterial infection that is of along duration or frequent recurrence. For example, a chronic middle earinfection, or otitis media, can occur when the Eustachian tube becomesblocked repeatedly due to allergies, multiple infections, ear trauma, orswelling of the adenoids. The definition of “long duration” will dependupon the particular infection. For example, in the case of a chronicmiddle ear infection, it may last for weeks to months. Other knownchronic bacterial infections include urinary tract infection (mostcommonly caused by Escherichia coli and/or Staphylococcussaprophyticus), gastritis (most commonly caused by Helicobacter pylori),respiratory infection (such as those commonly afflicting patents withcystic fibrosis, most commonly caused by Pseudomonas aeuroginosa),cystitis (most commonly caused by Escherichia coli), pyelonephritis(most commonly caused by Proteus species, Escherichia coli and/orPseudomonas species), osteomyelitis (most commonly caused byStaphylococcus aureus, but also by Escherichia coli), bacteremia, skininfection, rosacea, acne, chronic wound infection, infectious kidneystones (can be caused by Proteus mirabilis), bacterial endocarditis, andsinus infection. A common infection afflicting pigs is atrophic rhinitis(caused by Bordatella species, e.g. Bordatella rhinitis).

Also disclosed is a method of clearing a preformed biofilm from asubstrate comprising the step of administering an effective amount of acompound described herein to said substrate, wherein said effectiveamount will reduce the amount of said biofilm on said substrate.“Preformed biofilm” is a biofilm that has begun to adhere to asubstrate. The biofilm may or may not yet be fully formed.

Also provided are methods of treating subjects infected with abacterium. Methods of treating a subject infected with a bacterium cancomprise administering to the subject a therapeutically effective amountof a compound described herein. In some embodiments, the bacterium cancomprise a Gram-positive bacterium. For example, the bacterium caninclude Staphylococcus aureus (methicillin sensitive), Staphylococcusaureus (methicillin resistant), Streptococcus pneumonia (penicillinsensitive), Streptococcus pneumonia (penicillin resistant),Staphylococcus epidermis (multiple drug resistant), Enterococcusfaecalis (vancomycin sensitive), Enterococcus faecium (vancomycinresistant), and/or Haemophilus influenzae.

By way of non-limiting illustration, examples of certain embodiments ofthe present disclosure are given below.

EXAMPLES

Materials and Methods

THE was purified using an alumina filtration system. Aldehydes werepurchased from a commercial chemical company and used as received.Reactions were monitored by TLC analysis (pre-coated silica gel 60 F₂₅₄plates, 250 mm layer thickness) and visualization was accomplished witha 254 nm UV light and by staining with a KMnO₄ solution (1.5 g of KMnO₄,10 g of K2CO3, and 1.25 mL of a 10% NaOH solution in 200 mL of water).Reactions were also monitored by LC-MS (2.6 mm C18 50×2.10 mm column).Flash chromatography on SiO₂ was used to purify the crude reactionmixtures and performed on a flash system utilizing pre-packed cartridgesand linear gradients. Melting points were determined using a capillarymelting point apparatus. Infrared spectra were determined on a FT/IRspectrometer. ¹H and ¹³C NMR spectra were obtained on a 400 MHzinstrument in CDCl₃ unless otherwise noted. Chemical shifts werereported in parts per million with the residual solvent peak used as aninternal standard (CDCl₃=7.26 ppm for ¹H and 77.16 ppm for ¹³C). ¹H NMRspectra were run at 300, 400, or 700 MHz and are tabulated as follows:chemical shift, multiplicity (s=singlet, d=doublet, t=triplet,q=quartet, p=pentet, m=multiplet, bs=broad singlet, dt=doublet oftriplet, tt=triplet of triplet), number of protons, and couplingconstant(s). ¹³C NMR spectra were run at 100 or 175 MHz using aproton-decoupled pulse sequence with a di of 0 seconds unless otherwisenoted, and are tabulated by observed peak. High-resolution mass spectrawere obtained on an ion trap mass spectrometer using heated electrosprayionization (HESI).

General Procedure for the Synthesis of 2-Oxo-3-Propanoic Acids

2-oxo-3-propanoic acids were prepared according to establishedliterature procedures. In brief: A slurry of aldehyde (1 equiv.), sodiumacetate (1.3 equiv.) and N-acetylglycine (1.3 equiv.) in aceticanhydride (5 equiv.) was heated to 140° C. for 1 h. The reaction mixturewas quenched with ice (˜20 mL) with vigorous stirring and cooled in anice bath. The resulting precipitate was collected by vacuum filtrationto afford the azlactone, which was used without further purification.The azlactone was suspended in 3 M HCl and heated at reflux untilcomplete hydrolysis was observed by LC-MS (typically ˜3 h). The reactionmixture was cooled in an ice bath to facilitate crystallization. Theresulting solid was isolated by vacuum filtration and dried extensivelyunder high vacuum to provide the desired acid as a solid. This solid canbe recrystallized from benzene if necessary. Analytical data wasconsistent with previously reported data.

General Procedure for the Synthesis of α,α-Dichloroimines

α,α-dichloroimines were prepared according to an established literatureprocedure. In brief: To a solution of aldehyde in dry dichloromethane(0.2 M) was added dropwise t-butyl amine (1.0 equiv.) with vigorousstirring. To this solution was added anhydrous magnesium sulfate (˜3equiv.) and the mixture was stirred vigorously for ˜12 h at roomtemperature. The MgSO₄ was removed by vacuum filtration and the filtratewas concentrated in vacuo. The residue was dissolved in CCl₄ (0.5 M) andcooled to 0° C., before addition of N-chlorosuccinimide (2.1 equiv.) infour portions over the course of 2 hours. This mixture was stirred atroom temperature for 24 h. The mixture was cooled to 0° C. and filteredthrough a fine (4-5.5 μm) sintered glass frit funnel to removesuccinimide. The filtrate was concentrated in vacuo and driedextensively under high vacuum. The resulting α,α-dichloroimines wereused without purification and can be stored under inert atmosphere at−30° C.

Synthesis of 5-Benzylidene-4-Oxazolidinones

General Procedure: To a solution of arylpyruvic acid (˜0.5 mmol) in dryTHF (˜0.5 M) at 0° C. was added dropwise oxalyl chloride (1.15 equiv.)followed by N,N-dimethylformamide (0.10 equiv.). The solution wasstirred at 0° C. for 2 hours before the reaction mixture wasconcentrated under reduced pressure (directly through the septum using aneedle). The crude acid chloride was dissolved in dry CHCl₃ (0.5 M) andadded dropwise under inert atmosphere to a solution of crude t-butylα,α-dichloroimine in dry CHCl₃ (0.5 M) at 0° C. This mixture was warmedslowly to room temperature and stirred overnight. The reaction mixturewas concentrated in vacuo. The residue was dissolved in trifluoroaceticacid and heated to 50° C. until complete conversion was observed byLC-MS (typically 2-4 h). The reaction mixture was concentrated in vacuoand purified by reverse-phase flash chromatography (SiO₂—C18;CH₃CN/H₂O).

(Z)-2-(1,1-dichloropentyl)-5-(4-(trifluoromethyl)benzylidene)oxazolidin-4-one(1). 25 mg (22%) isolated as a white crystalline solid. H NMR: (400 MHz,CDCl₃) δ 8.93 (s, 1H), 7.74 (d, J=7.79 Hz, 2H), 7.64 (d, J=7.81 Hz, 2H),6.35 (s, 1H), 5.80 (s, 1H), 2.33 (m, 1H), 2.20 (m, 1H), 1.78 (m, 2H),1.46 (m, 2H), 0.99 (t, J=7.19, 3H).

(Z)-2-(1,1-dichloro-2-phenylethyl)-5-(4-(trifluoromethyl)benzylidene)oxazolidin-4-one(2). 18 mg (10%) isolated as a white crystalline solid. ¹H NMR: (400MHz, CDCl₃) δ 7.76 (d, J=8.35 Hz, 2H), 7.67 (d, J=8.39 Hz, 2H), 7.43 (m,2H), 7.38 (m, 3H), 6.38 (s, 1H), 5.71 (s, 1H), 3.69 (s, 2H).

(Z)-2-(1,1-dichlorononyl)-5-(4-(trifluoromethyl)benzylidene)oxazolidin-4-one(3). 23 mg (24%) isolated as an off-white solid. ¹H NMR: (400 MHz,CDCl₃) δ 8.94 (s, 1H), 7.74 (d, J=7.84 Hz, 2H), 7.64 (d, J=7.88 Hz, 2H),6.35 (s, 1H), 5.80 (s, 1H), 2.33 (m, 1H), 2.20 (m, 1H), 1.79 (m, 2H),1.41 (m, 2H), 1.35-1.29 (m, 8H), 0.99 (t, J=6.61, 3H).

(Z)-2-(1,1-dichloroethyl)-5-(4-(trifluoromethyl)benzylidene)oxazolidin-4-one(4). 18 mg (25%) isolated as a white crystalline solid. ¹H NMR: (400MHz, (CD₃)₂CO) δ 9.44 (s, 1H), 7.93 (d, J=8.07 Hz, 2H), 7.75 (d, J=7.76Hz, 2H), 6.28 (s, 1H), 6.07 (s, 1H), 2.28 (s, 3H).

(Z)-2-(1,1-dichlorobut-3-yn-1-yl)-5-(4-(trifluoromethyl)benzylidene)oxazolidin-4-one(5). 46 mg (51%) isolated as an off-white solid. ¹H NMR: (400 MHz,CDCl₃) δ 8.69 (s, 1H), 7.74 (d, J=7.85 Hz, 2H), 7.64 (d, J=7.86 Hz, 2H),6.38 (s, 1H), 6.06 (s, 1H), 3.37 (s, 2H), 2.34 (s, 1H).

(Z)-5-benzylidene-2-(1,1-dichloropentyl)oxazolidin-4-one (6). 19 mg(28%) isolated as a white solid. ¹H NMR: (400 MHz, CDCl₃) δ 8.83 (s,1H), 7.65 (d, J=7.19 Hz, 2H), 7.39 (m, 2H), 7.30 (m, 1H), 6.34 (s, 1H),5.77 (s, 1H), 2.32 (m, 1H), 2.20 (m, 1H), 1.77 (m, 2H), 1.44 (m, 2H),0.99 (t, J=7.34, 3H).

(Z)-2-(tert-butyl)-5-(4-(trifluoromethyl)benzylidene)oxazolidin-4-one(7). Synthesized via a previously reported procedure⁵. 46 mg (96%)isolated as a white solid. ¹H NMR: (400 MHz, CDCl₃) δ 9.02 (s, 1H), 7.74(d, J=8.31 Hz, 2H), 7.62 (d, J=8.33 Hz, 2H), 6.22 (s, 1H), 5.31 (s, 1H),1.04 (s, 9H).

(Z)-2-pentyl-5-(4-(trifluoromethyl)benzylidene)oxazolidin-4-one (8).Previously reported⁵. 8.7 mg (32%) isolated as a brown oil: ¹H NMR (400MHz, CDCl₃) δ 8.92 (s, 1H), 7.76 (d, 2H, J=8.0 Hz), 7.61 (d, 2H, J=8.1Hz), 6.26 (s, 1H), 5.71 (bs, 1H), 1.88 (s, 2H), 1.71-1.29 (m, 6H), 0.91(t, 3H, J=6.6 Hz); ¹³C NMR (100 MHz, CDCl₃) δ 165.8, 145.3, 137.1,129.3, 125.6, 122.9, 102.5, 89.7, 36.4, 31.5, 22.8, 22.6, 14.0. IR (thinfilm, cm⁻¹): 3213, 2930, 1714, 1324, 1117, 861; HRMS (HESI) m zcalculated for C₁₆H₁₉F₃NO₂ [M+H]⁺ 312.1217, found 312.1214; R_(f)=0.27(40% EtOAc/hexanes).

N-hexyl-2-oxo-3-(4-(trifluoromethyl)phenyl)propanamide (9). To asolution of 2-oxo-3-(4-(trifluoromethyl)phenyl)propanoic acid (0.232 g,1.0 mmol) in THE (10 mL; 0.1 M) at 0° C. was addedN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (0.2322 g,1.20 mmol, 1.2 equiv.), 1-hydroxybenzotriazole hydrate (0.1843 g, 1.20mmol, 1.2 equiv.) and N,N-diisopropylethylamine (0.355 mL, 2.0 mmol, 2.0equiv.). This mixture was stirred at 0° C. for several minutes beforehexylamine (0.146 mL, 1.0 mmol, 1.0 equiv.) was added and the resultingsolution was allowed to warm to room temperature with vigorous stirring.After 14 hours the reaction mixture was diluted with Et₂O and washedsuccessively with 3 M HCl (10 mL×2), sat. NaHCO₃ (10 mL×3), H₂O (20 mL)and brine. The organic layer was dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by flash chromatography(SiO₂, EtOAc/hexanes). 116 mg (43%) isolated as an off-white solid. ¹HNMR: (400 MHz, CDCl₃) δ 7.87 (d, J=7.82 Hz, 2H), 7.67 (d, J=7.81 Hz,2H), 4.58 (s, 2H), 3.60 (m, 2H), 2.33 (m, 1H), 2.20 (m, 1H), 1.83 (m,2H), 1.58 (m, 4H), 1.17 (m, 3H).

(±)-Synoxazolidinone A (10). As previously reported⁶: 10.9 mg (88%)isolated as a colorless oil: ¹H NMR (400 MHz, CD₃OD) δ 7.89 (s, 2H),6.08 (s, 1H), 5.91 (d, J=2.4 Hz, 1H), 4.30 (dt, J=11.0, 2.6 Hz, 1H),3.86 (s, 3H), 3.56-3.35 (m, 2H), 2.32-2.22 (m, 1H), 2.10-1.99 (m, 1H);¹³C NMR (100 MHz, CD₃OD) δ 165.6, 158.8, 154.5, 146.5, 134.1, 119.1,100.6, 90.3, 61.7, 61.2, 39.5, 32.3. HRMS (HESI) m z calculated forC₁₅H₁₆Br₂ClN₄O₃ [M−H] 492.9283, found 492.9290.

Biological Characterization of Compounds 1-10

Broth microdilution protocol for minimum inhibition concentration (MIC)determination. Fresh Mueller-Hinton broth (MHB) ˜ 3 ml, was inoculated(5×10⁵ CFU mL⁻¹) with MRSA (ATCC 33591) methicillin-resistantStaphylococcus aureus. The resulting bacterial suspension was aliquoted(1 mL) into culture tubes and test compound (10 mM DMSO stock solution)was added to reach the final concentration of the interest. Bacteria nottreated with the antimicrobial compound served as the control. Rows 2-12of a 96-well microtiter plate were filled with remaining bacterialsubcultures. The samples containing test compound were aliquoted (200μL) into the first row wells of the microtiter plate (2 wells percompound). Row 1 wells were mixed 6-8 times and 100 μL was transferreddown to row 2. Row 2 wells were mixed 6-8 times, followed by a 100 μLtransfer from row 2 down to row 3. The rest of the rows of the platewere serially diluted in similar fashion. The plate was covered withsealing tape for 96-well plates (ThermoFisher) and incubated understationary conditions at 37° C. A duplicate of the plate was prepared.The lowest concentration of test compound at which no visible growth ofbacteria occurred—minimum inhibitory concentration (MIC) values wererecorded after 16 h.

The results are included in Table 1 below.

Biofilm Inhibition Assay. Biofilm inhibition assays were performed bytaking an overnight culture of bacterial strain and subculturing it atan OD₆₀₀ of 0.01 into the necessary growth liquid medium (LB for A.baumannii, LBNS for P. aeruginosa, Stainer-Scholte medium that wassupplemented with 10 μL/mL of 100× nutrient complex for B.bronchiseptica and TSB w/0.3% glucose for S. aureus) for the strain. Thecompound being tested was then added at a predetermined concentrationand then aliquoted (100 μL) into the wells of a 96-well PVC microtiterplate (wells not used for samples are filled with 100 μL of de-ionizedwater). Plates were then wrapped in GLAD Press n' Seal and incubatedunder stationary conditions at 37° C. After 24 hours, the media wasdiscarded from the wells and the plates were washed thoroughly with tapwater. Plates were then stained with 100 μL of 0.1% solution of crystalviolet (CV) and then incubated at an ambient temperature for 30 minutes.Sample plates were then washed with tap water again and the remainingstain was solubilized with 200 μL of 95% ethanol. Biofilm inhibition wasquantitated by measuring the OD₅₄₀ for each well by transferring 125 μLof the solubilized CV stain into a polystyrene microtiter dish foranalysis.

The results are included in Table 1 below.

Biofilm Dispersion Assay. Dispersion assays were performed by taking anovernight culture of bacterial strain and subculturing it at an OD₆₀₀ of0.01 into the necessary growth liquid medium. The resulting bacterialsuspension was aliquoted (100 μL) into the wells of a 96-well PVCmicrotiter plate. Plates were then wrapped in GLAD Press n' Sealfollowed by an incubation under stationary conditions at an ambienttemperature. After 24 hours, the media was discarded from the wells andthe plates were washed thoroughly with tap water. Predeterminedconcentrations of the test compound were then made in the same mediumused to initially grow the biofilms and then aliquoted (100 μL) into thewells of the 96-well PVC microtiter plate with the established biofilms.Plates were then wrapped in GLAD Press n' Seal and incubated understationary conditions at 37° C. After 24 hours, the media was discardedfrom the wells and the plates were washed thoroughly with tap water.Plates were then stained with 100 μL of 0.1% solution of crystal violet(CV) and then incubated at room temperature for 30 minutes. Plates werethen washed with tap water again and the remaining stain was solubilizedwith 200 μL of 95% ethanol. Biofilm dispersion was quantitated bymeasuring the OD₅₄₀ for each well by transferring 125 μL of thesolubilized CV stain into a polystyrene microtiter dish for analysis.

The results are included in Table 1 below.

TABLE 1 Summary of the activity of Compounds 1-10. AntimicrobialActivity Biofilm Inhibition Biofilm Dispersion MIC (μg/mL) IC₅₀ (μM)IC₅₀ (μM) MRSA MRSA MRSA MRSA Compound ATCC 33591 ATCC BAA-44 ATCCBAA-44 ATCC BAA-44 1 12 12 1.0 5.7 2 6 6 1.4 4.3 3 >128 >128 1.7 6.6 432 32 3.6 5.0 5 16 16 1.9 6.0 6 64 64 3.2 5.3 7 128 128 7.3 9.2 8 16 167.3 ND 9 >128 >128 >20 >20 10 10 10 27 >20

The compounds, compositions, and methods of the appended claims are notlimited in scope by the specific compounds, compositions, and methodsdescribed herein, which are intended as illustrations of a few aspectsof the claims. Any compounds, compositions, and methods that arefunctionally equivalent are intended to fall within the scope of theclaims. Various modifications of the compounds, compositions, andmethods in addition to those shown and described herein are intended tofall within the scope of the appended claims. Further, while onlycertain representative compounds, compositions, and method stepsdisclosed herein are specifically described, other combinations of thecompounds, compositions, and method steps also are intended to fallwithin the scope of the appended claims, even if not specificallyrecited. Thus, a combination of steps, elements, components, orconstituents may be explicitly mentioned herein or less, however, othercombinations of steps, elements, components, and constituents areincluded, even though not explicitly stated.

The term “comprising” and variations thereof as used herein is usedsynonymously with the term “including” and variations thereof and areopen, non-limiting terms. Although the terms “comprising” and“including” have been used herein to describe various embodiments, theterms “consisting essentially of” and “consisting of” can be used inplace of “comprising” and “including” to provide for more specificembodiments of the invention and are also disclosed. Other than wherenoted, all numbers expressing geometries, dimensions, and so forth usedin the specification and claims are to be understood at the very least,and not as an attempt to limit the application of the doctrine ofequivalents to the scope of the claims, to be construed in light of thenumber of significant digits and ordinary rounding approaches.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meanings as commonly understood by one of skill in the artto which the disclosed invention belongs. Publications cited herein andthe materials for which they are cited are specifically incorporated byreference.

1. A compound defined by Formula I

or a pharmaceutically acceptable salt or prodrug thereof, wherein X ischosen from O and S; R¹, R², R³, R⁴, and R⁵ are each independentlychosen from hydrogen, halogen, hydroxyl, —CN, —NO₂, amino, alkylamino,dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy,haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl,haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl,haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁶ and R⁷ are each independently chosen fromhydrogen, hydroxy, halogen, —CN, —NO₂, amino, alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl, haloalkylsulfinyl,alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl,alkoxycarbonyl, haloalkoxycarbonyl, alkylaminocarbonyl,heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁸ is chosen from alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, alkylcycloheteroalkyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl, each optionallysubstituted with one or more substituents individually chosen from R¹¹;R⁹ is chosen from hydrogen, hydroxy, halogen, —CN, —NO₂, amino,alkylamino, dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio,alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, aryl,heteroaryl, cycloalkyl, cycloheteroalkyl, alkylaryl, alkylheteroaryl,alkylcycloalkyl, alkylcycloheteroalkyl, alkylsulfinyl,haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl,haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl, each optionally substituted with one or moresubstituents individually chosen from R¹¹; R¹⁰ is chosen from hydrogen,alkyl, haloalkyl, alkylthio, haloalkylthio, alkenyl, haloalkenyl,alkynyl, haloalkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, and alkylcycloheteroalkyl,each optionally substituted with one or more substituents individuallychosen from R¹¹; and R¹¹ is chosen from hydroxy, halogen, —CN, —NO₂,amino, alkylamino, dialkylamino, alkyl, haloalkyl; alkylthio;haloalkylthio; alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl; with the provisothat the compound is not one of the following:

2-23. (canceled)
 34. A method of controlling biofilm formation on asubstrate comprising contacting the substrate with a compound defined byFormula I

or a pharmaceutically acceptable salt or prodrug thereof in an amounteffective to inhibit biofilm formation, wherein X is chosen from O andS; R¹, R², R³, R⁴, and R⁵ are each independently chosen from hydrogen,halogen, hydroxyl, —CN, —NO₂, amino, alkylamino, dialkylamino, alkyl,haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl, haloalkylsulfinyl,alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl,alkoxycarbonyl, haloalkoxycarbonyl, alkylaminocarbonyl,heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁶ and R⁷ are each independently chosen fromhydrogen, hydroxy, halogen, —CN, —NO₂, amino, alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, haloalkynyl, alkylsulfinyl, haloalkylsulfinyl,alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl,alkoxycarbonyl, haloalkoxycarbonyl, alkylaminocarbonyl,heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁸ is chosen from alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, alkylcycloheteroalkyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl, each optionallysubstituted with one or more substituents individually chosen from R¹¹;R⁹ is chosen from hydrogen, hydroxy, halogen, —CN, —NO₂, amino,alkylamino, dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio,alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, aryl,heteroaryl, cycloalkyl, cycloheteroalkyl, alkylaryl, alkylheteroaryl,alkylcycloalkyl, alkylcycloheteroalkyl, alkylsulfinyl,haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl,haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl, each optionally substituted with one or moresubstituents individually chosen from R¹¹; R¹⁰ is chosen from hydrogen,alkyl, haloalkyl, alkylthio, haloalkylthio, alkenyl, haloalkenyl,alkynyl, haloalkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, and alkylcycloheteroalkyl,each optionally substituted with one or more substituents individuallychosen from R¹¹; and R¹¹ is chosen from hydroxy, halogen, —CN, —NO₂,amino, alkylamino, dialkylamino, alkyl, haloalkyl; alkylthio;haloalkylthio; alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl.
 35. The method ofclaim 34, wherein the compound defined by Formula I comprises a compounddefined by claim
 1. 36. The method of claim 34, wherein the biofilmcomprises Gram-positive bacteria.
 37. The method of claim 36, whereinthe biofilm comprises bacteria of a genus Staphylococcus.
 38. The methodof claim 37, wherein the biofilm comprises bacteria of the speciesStaphylococcus aureus.
 39. A method for treating a chronic bacterialinfection in a subject in need thereof, comprising administering to saidsubject a compound defined by Formula I

or a pharmaceutically acceptable salt or prodrug thereof in an amounteffective to inhibit, reduce, or remove a biofilm component of thechronic bacterial infection, wherein X is chosen from O and S; R¹, R²,R³, R⁴, and R⁵ are each independently chosen from hydrogen, halogen,hydroxyl, —CN, —NO₂, amino, alkylamino, dialkylamino, alkyl, haloalkyl,alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl, haloalkenyl,alkynyl, haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl; R⁶ and R⁷ are eachindependently chosen from hydrogen, hydroxy, halogen, —CN, —NO₂, amino,alkylamino, dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio,alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl,alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl,alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl; R⁸ is chosen from alkylamino, dialkylamino,alkyl, haloalkyl, alkylthio, haloalkylthio, alkoxy, haloalkoxy, alkenyl,haloalkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, alkylcycloheteroalkyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl, each optionallysubstituted with one or more substituents individually chosen from R¹¹;R⁹ is chosen from hydrogen, hydroxy, halogen, —CN, —NO₂, amino,alkylamino, dialkylamino, alkyl, haloalkyl, alkylthio, haloalkylthio,alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl, haloalkynyl, aryl,heteroaryl, cycloalkyl, cycloheteroalkyl, alkylaryl, alkylheteroaryl,alkylcycloalkyl, alkylcycloheteroalkyl, alkylsulfinyl,haloalkylsulfinyl, alkylsulfonyl, haloalkylsulfonyl, alkylcarbonyl,haloalkylcarbonyl, alkoxycarbonyl, haloalkoxycarbonyl,alkylaminocarbonyl, heteroalkylaminocarbonyl, dialkylaminocarbonyl, andheterodialkylaminocarbonyl, each optionally substituted with one or moresubstituents individually chosen from R¹¹; R¹⁰ is chosen from hydrogen,alkyl, haloalkyl, alkylthio, haloalkylthio, alkenyl, haloalkenyl,alkynyl, haloalkynyl, aryl, heteroaryl, cycloalkyl, cycloheteroalkyl,alkylaryl, alkylheteroaryl, alkylcycloalkyl, and alkylcycloheteroalkyl,each optionally substituted with one or more substituents individuallychosen from R¹¹; and R¹¹ is chosen from hydroxy, halogen, —CN, —NO₂,amino, alkylamino, dialkylamino, alkyl, haloalkyl; alkylthio;haloalkylthio; alkoxy, haloalkoxy, alkenyl, haloalkenyl, alkynyl,haloalkynyl, alkylsulfinyl, haloalkylsulfinyl, alkylsulfonyl,haloalkylsulfonyl, alkylcarbonyl, haloalkylcarbonyl, alkoxycarbonyl,haloalkoxycarbonyl, alkylaminocarbonyl, heteroalkylaminocarbonyl,dialkylaminocarbonyl, and heterodialkylaminocarbonyl.
 40. The method ofclaim 39, wherein the compound defined by Formula I comprises a compounddefined by claim
 1. 41. The method of any of claims 39-40, wherein thechronic bacterial infection is chosen from urinary tract infection,gastritis, respiratory infection, cystitis, pyelonephritis,osteomyelitis, bacteremia, skin infection, rosacea, acne, chronic woundinfection, infectious kidney stones, bacterial endocarditis, and sinusinfection. 42-48. (canceled)